Anti-methanogenic potential of seaweeds and seaweed-derived compounds in ruminant feed:current perspectives,risks and future prospects

With methane emissions from ruminant agriculture contributing 17%of total methane emissions worldwide,there is increasing urgency to develop strategies to reduce greenhouse gas emissions in this sector.One of the proposed strategies is ruminant feed intervention studies focused on the inclusion of anti-methanogenic compounds which are those capable of interacting with the rumen microbiome,reducing the capacity of ruminal microorganisms to produce methane.Recently,seaweeds have been investigated for their ability to reduce methane in ruminants in vitro and in vivo,with the greatest methane abatement reported when using the red seaweed Asparagopsis taxiformis(attributed to the bromoform content of this species).From the literature analysis in this study,levels of up to 99%reduction in ruminant methane emissions have been reported from inclusion of this seaweed in animal feed,although further in vivo and microbiome studies are required to confirm these results as other reports showed no effect on methane emission resulting from the inclusion of seaweed to basal feed.This review explores the current state of research aiming to integrate seaweeds as anti-methanogenic feed additives,as well as examining the specific bioactive compounds within seaweeds that are likely to be related to these effects.The effects of the inclusion of seaweeds on the ruminal microbiome are also reviewed,as well as the future challenges when considering the largescale inclusion of seaweeds into ruminant diets as anti-methanogenic agents.

Limosilactobacillus mucosae FZJTZ26M3 prevents NAFLD in mice through modulation of lipid metabolism and gut microbiota dysbiosis

Lactobacillus are considered promising therapeutic methods for nonalcoholic fatty liver disease(NAFLD).The effects of two strains of Ltmosilactobacillus mucosae on NAFLD were investigated in this study.Fourweek-old male C57BL/6J mice were divided into 4 groups(n=8 per group,Control,Model,FZJTZ26M3,FGSYC17L3).L.mucosae FZJTZ26M3 reduced the mice 's body weight,liver weight,and adipose tissue weight after 12 weeks of therapy.According to serum analysis,total cholesterol,triacylglycerol,and low-density lipoprotein cholesterol significantly decreased after L.mucosae FZJTZ26M3 intervention.Liver pathology showed that L.mucosae FZJTZ26M3 was effective to ameliorate lipid deposition in NAFLD mice.Additionally,the expression of the gene related to lipid metabolism in the liver and adipose tissue was analyzed,and the results indicated that L.mucosae FZJTZ26M3 could alleviate NAFLD by regulating lipid metabolism.Furthermore,the results of 16S rRNA gene sequencing revealed a drop in the relative abundance of Ruminococcaceae,which is linked to inflammation,but the relative abundance of a potential probiotic Akkermansia significantly increased after L.mucosae FZJTZ26M3 intervention.Generally,L.mucosae FZJTZ26M3 could be a candidate to prevent NAFLD.

Structural and Functional Annotation of Hypothetical Protein of Fusobacterium nucleatum Strain MJR7757B: An in Silico Approach

Fusobacterium nucleatum is an anaerobic, commensal, gram-negative oral bacterium that is carcinogenic and causes a wide range of human diseases. The present study focused on the analysis of the hypothetical protein, HMPREF3221_01179, derived from F. nucleatum strain MJR7757B, employing various computational methods to anticipate both its structure and functional characteristics. NCBI conserved domain analysis, NCBI BLASTp and MEGA Phylogenetic tree study characterize the target protein as an outer membrane efflux protein (ToIC family) which facilitate the bacterial transmembrane transport. With a molecular weight of 52120.02 Da, an isoelectric point (pI) of 8.33, and an instability index of 29.47, the protein is anticipated to exhibit good solubility in the extracellular space and crucial stability for pharmaceutical applications. The protein’s structure meets quality standards during the construction and refinement of its 3D model. The efflux inhibitor Arginine beta-naphthylamide exhibits a significant binding affinity (-7.1 kcal/mol) to the binding site of the target protein. The in-silico analysis improves the understanding of the protein and facilitates future investigations into therapeutic medication.

Preserved egg white alleviates DSS-induced colitis in mice through the reduction of oxidative stress,modulation of inflammatory cytokines,NF-κB,MAPK and gut microbiota composition

Traditional Chinese preserved egg products have exhibited some anti-inflammatory effects,but their mechanisms of action remain unknown.This study aimed to investigate the anti-inflammatory effects of preserved egg white(PEW)treatment on dextran sulfate sodium(DSS)-induced colitis in mice and the underlying mechanisms.The results showed that treatment with PEW in mice with DSS-induced colitis for 14 days effectively improved the clinical signs,inhibited the secretion and gene expression of pro-inflammatory cytokines,and reduced myeloperoxidase(MPO)activity and oxidative stress levels.In addition,western blotting results showed that PEW significantly suppressed DSS-induced phosphorylation levels of nuclear factor-kappa B(NF-κB)p65 and p38 mitogen-activated protein kinase(MAPK)in colon tissues of mice with colitis.PEW also enhanced the production of short-chain fatty acids(SCFAs)and modulated gut microbiota composition in mice with DSS-induced colitis,including increasing the relative abundance of beneficial bacteria Lachnospiraceae,Ruminococcaceae and Muribaculaceae,and reducing the relative abundance of harmful bacteria Proteobacteria.Taken together,our study demonstrated that preserved egg white could alleviate DSS-induced colitis in mice through the reduction of oxidative stress,modulation of inflammatory cytokines,NF-κB,MAPK and gut microbiota composition.

Structural and antioxidative properties of royal jelly protein by partial enzymatic hydrolysis

The objective of this study was to investigate the structural and antioxidative properties of royal jelly protein(RJP)at different degrees of hydrolysis(DH)by partial enzymatic hydrolysis. RJP was hydrolyzed by alcalase for 0 min, 15 min, 1 h, 5 h and 8 h to obtain hydrolysates at DH of 5.34%, 11.65%, 15.19%, 21.38% and 23.91%, respectively. With the increased DH, the RJP hydrolysates showed elevated antioxidative activities. The molecular weight of RJP hydrolysates was significantly decreased but their primary backbone kept unchanged. Analysis of circular dichroism spectra revealed that the enzymolysis reduced the content of α-helix but increased the contents of β-sheet, β-turn and random coil. Meanwhile, the surface hydrophobicity and fluorescence intensity of RJP hydrolysates were decreased and a red shift occurred. As the enzymolysis continued, the surface morphology of RJP was gradually changed from a sheet-like structure into microparticles. Changes in antioxidative activities and structures generally followed a DH-dependent manner, however these changes became insignificant for samples at DH beyond 20%. Taking into consideration of both effectiveness and productivity, the optimum enzymatic duration was determined at 5 h.

Formation mechanisms of ethyl acetate and organic acids in Kluyveromyces marxianus L1-1 in Chinese acid rice soup

This study aims to explore the formation mechanism of ethyl acetate and organic acids in acid rice soup(rice-acid soup)inoculated with Kluyveromyces marxianus L1-1 through the complementary analysis of transcriptome and proteome.The quantity of K.marxianus L1-1 varied significantly in the fermentation process of rice-acid soup and the first and third days were the two key turning points in the growth phase of K.marxianus L1-1.Importantly,the concentrations of ethyl acetate,ethanol,acetic acid,and L-lactic acid increased from day 1 to day 3.At least 4231 genes and 2937 proteins were identified and 610 differentially expressed proteins were annotated to 30 Kyoto Encyclopedia of Genes and Genomes(KEGG)pathways based on the analysis results of transcriptome and proteome.The key genes and proteins including up-regulated alcohol dehydrogenase family,alcohol O-acetyltransferase,acetyl-CoA C-acetyltransferase,acyl-coenzyme A thioester hydrolase,and down-regulated aldehyde dehydrogenase family were involved in glycolysis/gluconeogenesis pathways,starch and sucrose metabolism pathways,amino sugar and nucleotide sugar metabolism pathways,tricarboxylic acid(TCA)cycle,and pyruvate metabolism pathways,thus promoting the formation of ethyl acetate,organic acids,alcohols,and other esters.Our results revealed the formation mechanisms of ethyl acetate and organic acids in rice-acid soup inoculated with K.marxianus L1-1.

Integrative proteomic-transcriptomic analysis revealed the lifestyles of Lactobacillus paracasei H4-11 and Kluyveromyces marxianus L1-1 under co-cultivation conditions

Compared with the rice-acid soup inoculated with single starter,the synergistically intensifi ed rice-acid soup inoculated with Lactobacillus paracasei H4-11(L.paracasei H4-11)and Kluyveromyces marxianus L1-1(K.marxianus L1-1)contained more fl avor compounds.Organic acids mainly included L-lactic acid and the main volatile fl avor component was ethyl acetate.Moreover,the signal intensity of astringency and bitterness and the total concentration of volatile sulfur compounds were reduced.The combined analysis results of RNA sequencing(RNA-seq)technology and 4D label-free quantitative(4D LFQ)proteomics explained the fl avor formation pathways in rice-acid soup inoculated with L.paracasei H4-11 and K.marxianus L1-1.In L.paracasei H4-11,L-lactate dehydrogenase,phosphoglucomutase,acetate kinase,alcohol dehydrogenase and acetyl-CoA were up-regulated and D-lactate dehydrogenase and N-Acetyltransferase were down-regulated.In K.marxianus L1-1,Acetyl-CoA,acetaldehyde dehydrogenase,acyl-coenzyme A,N-acetyltransferase,and L-lactate dehydrogenase were up-regulated and hexokinase,alcohol dehydrogenase,and alcohol O-acetyltransferase were down-regulated.The above up-regulation and down-regulation synergistically promoted the formation of characteristic fl avor compounds(mainly L-lactic acid and ethyl acetate).Enzyme-linked immunoassay(ELISA)and parallel reaction monitoring(PRM)quantitative analysis respectively verifi ed that 5 key metabolic enzymes and 27 proteins in L.paracasei H4-11 and K.marxianus L1-1 were associated with the characteristic fl avor of rice-acid soup,as confi rmed by the quantitative results of 4D LFQ.

Anti-lipid-oxidation effects and edible safety evaluation of the oil extracted by a supercritical CO_(2) process from coix seed fermented by Monascus purpureus

The physicochemical properties and composition of coix seed oil produced by Monascus purpureus fermentation and supercritical CO_(2)extraction were determined.Anti-lipid-oxidation and edible safety were evaluated using a cholesterol-fish oil model,acute oral toxicity assay,and genetic toxicity assay in vitro and in vivo,respectively.The results show that the extraction oil from fermented coix seed(FCS-O)had good physicochemical quality and abundant active components with physiological function.In particular,γ-tocotrienol,γ-oryzanol,coixenolide and oleic acid concentrations reached 72.83μg/g,745.96μg/g,9.65 mg/g and 316.58 mg/100 g DW,respectively.The FCS-O exhibited higher antioxidant capability in inhibiting lipid oxidation and peroxidation.Compared to the blank control,the concentrations of 7-ketocholestreol and peroxide only were 8.42μg/mL and 16.16 mmol/kg at 168 h of oxidation(P<0.01).In addition,the FCS-O has been confirmed to be a very safe edible oil,with no acute toxicity(LD50>10 g/kg bw,considered actually non-toxic)and no induced mutagenicity,cytotoxicity or genotoxicity.These results serve as a good safety reference for future application of the oil from fermented coix seed.The development and utilization of this kind of oil will be beneficial as a food,food ingredient,nutritional supplement,or natural food antioxidant to promote good health function.

Novel,non-colonizing,single-strain live biotherapeutic product ADS024 protects against Clostridioides difficile infection challenge in vivo

BACKGROUND The Centers for Disease Control and Prevention estimate that Clostridioides difficile(C.difficile)causes half a million infections(CDI)annually and is a major cause of total infectious disease death in the United States,causing inflammation of the colon and potentially deadly diarrhea.We recently reported the isolation of ADS024,a Bacillus velezensis(B.velezensis)strain,which demonstrated direct in vitro bactericidal activity against C.difficile,with minimal collateral impact on other members of the gut microbiota.In this study,we hypothesized that in vitro activities of ADS024 will translate in vivo to protect against CDI challenge in mouse models.AIM To investigate the in vivo efficacy of B.velezensis ADS024 in protecting against CDI challenge in mouse models.METHODS To mimic disruption of the gut microbiota,the mice were exposed to vancomycin prior to dosing with ADS024.For the mouse single-dose study,the recovery of ADS024 was assessed via microbiological analysis of intestinal and fecal samples at 4 h,8 h,and 24 h after a single oral dose of 5×108 colony-forming units(CFU)/mouse of freshly grown ADS024.The single-dose study in miniature swine included groups that had been pre-dosed with vancomycin and that had been exposed to a dose range of ADS024,and a group that was not pre-dosed with vancomycin and received a single dose of ADS024.The ADS024 colonies[assessed by quantitative polymerase chain reaction(qPCR)using ADS024-specific primers]were counted on agar plates.For the 28-d miniature swine study,qPCR was used to measure ADS024 levels from fecal samples after oral administration of ADS024 capsules containing 5×109 CFU for 28 consecutive days,followed by MiSeq compositional sequencing and bioinformatic analyses to measure the impact of ADS024 on microbiota.Two studies were performed to determine the efficacy of ADS024 in a mouse model of CDI:Study 1 to determine the effects of fresh ADS024 culture and ADS024 spore preparations on the clinical manifestations of CDI in mice,and Study 2 to compare the efficacy of single daily doses vs dosing 3 times per day with fresh ADS024.C.difficile challenge was performed 24 h after the start of ADS024 exposure.To model the human distal colon,an anerobic fecal fermentation system was used.MiSeq compositional sequencing and bioinformatic analyses were performed to measure microbiota diversity changes following ADS024 treatment.To assess the potential of ADS024 to be a source of antibiotic resistance,its susceptibility to 18 different antibiotics was tested.RESULTS In a mouse model of CDI challenge,single daily doses of ADS024 were as efficacious as multiple daily doses in protecting against subsequent challenge by C.difficile pathogen-induced disease.ADS024 showed no evidence of colonization based on the observation that the ADS024 colonies were not recovered 24 h after single doses in mice or 72 h after single doses in miniature swine.In a 28-d repeat-dose study in miniature swine,ADS024 was not detected in fecal samples using plating and qPCR methods.Phylogenetic analysis performed in the human distal colon model showed that ADS024 had a selective impact on the healthy human colonic microbiota,similarly to the in vivo studies performed in miniature swine.Safety assessments indicated that ADS024 was susceptible to all the antibiotics tested,while in silico testing revealed a low potential for off-target activity or virulence and antibioticresistance mechanisms.CONCLUSION Our findings,demonstrating in vivo efficacy of ADS024 in protecting against CDI challenge in mouse models,support the use of ADS024 in preventing recurrent CDI following standard antibiotic treatment.

瘤胃微生物对反刍动物饲料效率和甲烷排放的影响及其营养调控研究进展

饲料效率提高和甲烷排放的减少是反刍动物养殖的最主要目标之一,而瘤胃微生物对反刍动物的生产效率和健康状况以及温室气体排放起着至关重要的影响,但直到随着微生物多组学、高精仪器分析和生物信息技术的结合应用,才逐步建立了饲料效率、甲烷排放与瘤胃微生物之间的关系,从而更好地增加了对瘤胃微生物基因功能的理解。本文综述了瘤胃微生物组成与反刍动物饲料效率、甲烷排放的相关性及其营养调控的最新研究成果,旨在为进一步调节瘤胃微生物来提高反刍动物饲料效率和减少甲烷排放提供参考依据,也为克服这些障碍指出了潜在的营养途径。

PET分子量对PET/PC共混体系高压结晶行为的影响

以往高压合成的聚合物伸直链晶体尺寸较小且合成时间过长,不能满足单晶应用的要求。加入10wt%PC于不同分子量的PET中,采用自制高压装置,制备共混体系,用WAXD,SEM,DSC等表征手段,研究了PET/PC共混体系在高压下的结晶行为。研究结果表明:通过PET/PC体系在高压下化学反应诱导的自组装,不同分子量PET的PET/PC共混体系均在较短的时间内生成了尺寸较大的聚酯伸直链晶体,对应于三斜晶系,其中PET/PC体系最适于大尺寸伸直链晶体的合成,仅需6hr时间即生成PE需200hr方可合成的40μm厚度的伸直链晶体。同时提出了PET1/PC体系中伸直链晶体高温下缺陷自我修复模型。

Biosorption characteristics of unicellular green alga Chlorella sorokiniana immobilized in loofa sponge for removal of Cr(Ⅲ)

Loofa sponge （LS） immobilized biomass of Chlorella sorokiniana （LSIBCS）, isolated from industrial wastewater, was investigated as a new biosorbent for the removal of Cr（Ⅲ） from aqueous solution. A comparison of the biosorption of Cr（Ⅲ） by LSIBCS and free biomass of C. sorokiniana （FBCS） from 10-300 mg Cr（Ⅲ）/L aqueous solutions showed an increase in uptake of 17.79% when the microalgal biomass was immobilized onto loofa sponge. Maximum biosorption capacity for LSIBCS and FBCS was found to be 69.26 and 58.80 mg Cr（Ⅲ）/g biosorbent, respectively, whereas the amount of Cr（Ⅲ） ions adsorbed onto naked LS was 4.97 mg/g. The kinetics of Cr（Ⅲ） biosorption was extremely rapid and equilibrium was established in about 15 and 20 min by LSIBCS and FBCS, respectively. The biosorption equilibrium was well defined by Langmuir adsorption isotherm model. The biosorption kinetics followed the pseudo-second order kinetic model. The biosorption was found to be pH dependent and the maximum sorption occurred at the solution pH 4.0. Desorption studies showed that 98% of the adsorbed Cr（Ⅲ） could be desorbed with 0.1 mol/L HNO3, while other desorbing agents were less effective in the order： EDTA 〉 H2SO4 〉 CH3COOH 〉 HCl. The regenerated LSIBCS retained 92.68% of the initial Cr（Ⅲ） binding capacity up to five cycles of reuse in continuous flow-fixed bed columns. The study revealed that LSIBCS could be used as an effective biosorbent for the removal of Cr（Ⅲ） from wastewater.

Bemisia tabaci Phylogenetic Groups in India and the Relative Transmission Efficacy of Tomato leaf curl Bangalore virus by an Indigenous and an Exotic Population

Bemisia tabaci adults from various host-plant species were collected from 31 regions across India.266 B.tabaci samples were first screened by RAPD-PCR to examine molecular variability and to select individuals with different fingerprints.Host-plant and region of collection were then used to select 25 individuals for PCR amplification and sequencing of their partial mitochondrial cytochrome oxidase subunit one（mtCOI）genes.Pairwise comparisons with mtCOI consensus sequences showed that the majority of these samples had 〈3.5% sequence divergence from groups currently termed Asia I,Asia II-5,Asia II-7,and Asia II-8.The biotype-B B.tabaci from India clustered into the Middle East-Asia Minor 1 group.A new group of B.tabaci from Coimbatore,collected from pumpkin,was related most closely to the Asia I group（6.2% sequence divergence from the consensus Asia I sequence）.To increase our understanding of the epidemiology of tomato leaf curl disease（ToLCD）and the key B.tabaci genetic groups involved in virus spread,the indigenous Asia I and the exotic biotype-B population from South India were used to carry out transmission experiments using Tomato leaf curl Bangalore virus（ToLCBV）.The acquisition access periods（AAP）,inoculation access periods（IAP）,latent periods（LP）,and ToLCBV transmission efficiencies of the two populations were compared and the biotype-B had the more efficient transmission characteristics.These results are discussed in relation to recent changes in the epidemiology of tomato leaf curl disease in South India.

Changes in the colon microbiota and intestinal cytokine gene expression following minimal intestinal surgery

AIM: To investigate the impact of minor abdominal surgery on the caecal microbial population and on markers of gut inflammation.METHODS: Four week old piglets were randomly allocated to a no-surgery "control" group(n = 6) or a "transection surgery" group(n = 5).During the transection surgery procedure, a conventional midline incision of the lower abdominal wall was made and the small intestine was transected at a site 225 cm proximal to the ileocaecal valve, a 2 cm segment was removed and the intestine was re-anastomosed.Piglets received a polymeric infant formula diet throughout the study period and were sacrificed at two weeks post-surgery.Clinical outcomes including weight, stool consistency and presence of stool fat globules were monitored.High throughput DNA sequencing of colonic content was used to detect surgery-relateddisturbances in microbial composition at phylum, family and genus level.Diversity and richness estimates were calculated for the control and minor surgery groups.As disturbances in the gut microbial community are linked to inflammation we compared the gene expression of key inflammatory cytokines(TNF, IL1 B, IL18, IL12, IL8, IL6 and IL10) in ileum, terminal ileum and colon mucosal extracts obtained from control and abdominal surgery groups at two weeks post-surgery.RESULTS: Changes in the relative abundance of bacterial species at family and genus level were confined to bacterial members of the Proteobacteria and Bacteroidetes phyla.Family level compositional shifts included a reduction in the relative abundance of Enterobacteriaceae(22.95 ± 5.27 vs 2.07 ± 0.72, P < 0.01), Bacteroidaceae(2.54 ± 0.56 vs 0.86 ± 0.43, P < 0.05) and Rhodospirillaceae(0.40 ± 0.14 vs 0.00 ± 0.00, P < 0.05) following transection surgery.Similarly, at the genus level, changes associated with transection surgery were restricted to members of the Proteobacteria and Bacteroidetes phyla and included decreased relative abundance of Enterobacteriaceae(29.20 ± 6.74 vs 2.88 ± 1.08, P < 0.01), Alistipes(4.82 ± 1.73 vs 0.18 ± 0.13, P < 0.05) and Thalassospira(0.53 ± 0.19 vs 0.00 ± 0.00, P < 0.05).Surgeryassociated microbial dysbiosis was accompanied by increased gene expression of markers of inflammation.Within the ileum IL6 expression was decreased(4.46 ± 1.60 vs 0.24 ± 0.06, P < 0.05) following transection surgery.In the terminal ileum, gene expression of TNF was decreased(1.51 ± 0.13 vs 0.80 ± 0.16, P < 0.01) and IL18(1.21 ± 0.18 vs 2.13 ± 0.24, P < 0.01), IL12(1.04 ± 0.16 vs 1.82 ± 0.32, P < 0.05) and IL10(1.04 ± 0.06 vs 1.43 ± 0.09, P < 0.01) gene expression increased following transection surgery.Within the colon, IL12(0.72 ± 0.13 vs 1.78 ± 0.28, P < 0.01) and IL10(0.98 ± 0.02 vs 1.95 ± 0.14, P < 0.01) gene expression were increased following transection surgery.CONCLUSION: This study suggests that minor abdominal surgery in infants, results in long-term alteration of the colonic microbial composition and persistent gastrointestinal inflammation.

Biodegrading Wheat Bran with Agaricus blazei and Its Effects on Intestinal Development Identified with Mice

Agaricus blazei Murill is a well known edible/medicinal mushroom used for immune-nutrient food therapies.The biodegradation of different substrates with this fungus may result in different metabolites and degraded compounds,which may enrich the function of the food therapies.In this study,soluble compounds from the culture of A.blazei grown in liquid media with whole wheat bran and its water filtrate,respectively,were compared.Total soluble sugar,arabinoxylan,protein,and amino acids were significantly higher in the fungal culture resulted from the medium with whole wheat bran （43.54,1.56,0.59,and 2.19 mg mL^-1,respectively） than that from the medium with the bran filtrate （17.28,0.37,0.13,and 1.13 mg mL^-1,respectively）.The biodegraded wheat bran with cultured mycelia was fed to Mus musculus Linnaeus as supplementation or as dietary fiber exclusive ingredient.As non-specific food therapies,feeding effects on mice intestinal development were indicated indirectly in growth performance,intestinal absorption and serum parameters.Compared to feeding uncultured wheat bran,feeding mice with culture mixes of A.blazei resulted in remarkably increase in villus height and villus height/crypt depth ratio,which were increased by 25.4 and 31.0%,respectively,when applied as supplementation,and by 44.3 and 43.4%,respectively,when applied as dietary fiber.These increases are concomitant with the higher level of D-xylose in blood serum about 16.9 and 29.2% as supplementation and dietary fiber,respectively.The results implying that culturing A.blazei with whole wheat bran enhanced extracellular metabolism of the fungus and extensive degradation of wheat bran insoluble fibrous compounds.Furthermore,feeding the culture mix including metabolites and degraded wheat bran improved intestinal villus development,proving the nutritional benefits of the A.blazei mycelial cultures.

Characteristics of traditional Chinese acidic rice soup(rice-acid)prepared with different fermentation methods

Rice-acid,a Chinese traditional acidic rice soup(rice-acid),is widely accepted by consumers due to its unique flavor and anti-oxidation,anti-aging and immunity enhancement functions.This study confirmed that L-lactic acid and malic acid were the main organic acids in rice-acid.Low-temperature rice-acid samples produced by enterprises had the highest signal intensity of sour taste.The total content of free amino acids in different fermented rice-acid samples were in the range of 0.003-0.468 mg/g.42 key volatile flavor compounds were identified in rice-acid.8 volatile compounds with a higher contribution to the aroma of rice-acid were respectively acetic acid,1-octen-3-ol,2-heptanol,ethyl acetate,propyl propionate,hexanal,nonanal,and2,3-butanedione.The interaction between lactic acid bacteria(3.00×10^(3)-7.02×10^(6) CFU/mL)and yeasts(5.04×10^(4)-2.25×10^(8) CFU/mL)affected the formation of taste and aroma components in rice-acid.The physicochemical characteristics including titratable acidity,pH,reducing sugars,amino acid nitrogen,gammaaminobutyric acid showed significant differences between low-temperature fermentation samples and hightemperature fermentation samples.In addition,relationships linking all data through Pearson coefficient correlation were also reported.In summary,the study can be used to improve the quality of rice-acid products.

Rapid Profiling of Alkaloids in Several Medicinal Herbs by Matrix-assisted Laser Desorption/Ionization Mass Spectrometry

A simple method was developed for rapid and direct profiling of alkaloids in medical herbs via matrix-assisted laser desorption/ionization time-of-flight mass spectrometry（MALDI-TOF MS）. The dry herbs were first ground to powder and passed through a stainless steel sieve, mixed with the matrix solution to form a homogeneous suspension, which was then directly applied to MALDI analysis. Several matrices were investigated and 2,5- dihydroxybenzoic acid（DHB） was chosen as the optimized one, and the particle with small size was found to favor the analysis. Using this method, the profiles of alkaloids in several medical herbs were readily obtained, and the toxicities of crude and processed Radix Aconiti Lateralis Preparata were compared via the relative intensities of the peaks of the corresponding toxic components shown in their MALDI spectra. This method therefore provides a rapid and reliable protocol for obtaining profiles of alkaloids in medical herbs by using MALDI-TOF MS.

Genetic and genomic resources for Rubus breeding:a roadmap for the future

Rubus fruits are high-value crops that are sought after by consumers for their flavor,visual appeal,and health benefits.To meet this demand,production of red and black raspberries(R.idaeus L.and R.occidentalis L.),blackberries(R.subgenus Rubus),and hybrids,such as Boysenberry and marionberry,is growing worldwide.Rubus breeding programmes are continually striving to improve flavor,texture,machine harvestability,and yield,provide pest and disease resistance,improve storage and processing properties,and optimize fruits and plants for different production and harvest systems.Breeders face numerous challenges,such as polyploidy,the lack of genetic diversity in many of the elite cultivars,and until recently,the relative shortage of genetic and genomic resources available for Rubus.This review will highlight the development of continually improving genetic maps,the identification of Quantitative Trait Loci(QTL)s controlling key traits,draft genomes for red and black raspberry,and efforts to improve gene models.The development of genetic maps and markers,the molecular characterization of wild species and germplasm,and highthroughput genotyping platforms will expedite breeding of improved cultivars.Fully sequenced genomes and accurate gene models facilitate identification of genes underlying traits of interest and enable gene editing technologies such as CRISPR/Cas9.

Chromosome-scale scaffolding of the black raspberry (Rubus occidentalis L.) genome based on chromatin interaction data

Black raspberry(Rubus occidentalis L.)is a niche fruit crop valued for its flavor and potential health benefits.The improvement of fruit and cane characteristics via molecular breeding technologies has been hindered by the lack of a high-quality reference genome.The recently released draft genome for black raspberry(ORUS 4115-3)lacks assembly of scaffolds to chromosome scale.We used high-throughput chromatin conformation capture(Hi-C)and Proximity-Guided Assembly(PGA)to cluster and order 9650 out of 11,936 contigs of this draft genome assembly into seven pseudo-chromosomes.The seven pseudo-chromosomes cover~97.2%of the total contig length(~223.8 Mb).Locating existing genetic markers on the physical map resolved multiple discrepancies in marker order on the genetic map.Centromeric regions were inferred from recombination frequencies of genetic markers,alignment of 303 bp centromeric sequence with the PGA,and heat map showing the physical contact matrix over the entire genome.We demonstrate a high degree of synteny between each of the seven chromosomes of black raspberry and a high-quality reference genome for strawberry(Fragaria vesca L.)assembled using only PacBio long-read sequences.We conclude that PGA is a cost-effective and rapid method of generating chromosome-scale assemblies from Illumina short-read sequencing data.

Bacteriophage Biocontrol Rescues Mice Bacteremic of Clinically Isolated Mastitis from Dairy Cows Associated with Methicillin-Resistant <i>Staphyloccocus aureus</i>

Methicillin-resistant Staphylococcus aureus (MRSA) is among the most alarming pathogens affecting both humans and the global bovine industry. The current control measures in hospitals and on farms for MRSA have proven to be inadequate leaving a need for new rapid control methods to curb MRSA infections in situ. New control measures for bacterial infection are widely sought, with particular interest in the applications for bacteriophages (phages) as a biocontrol or therapeutic agent. The current study uses a wild highly lytic phage isolated from cow’s milk taken from three farms in Baghdad, Iraq. The resulting phage was able to rescue 100% of the mice from a median lethal dose (LD50) or (1 × 108 CFU mL-1 per mouse) for MRSA wild isolates achieved when the phage: bacteria ratio was 100:1. Even when treatment was delayed for 6 h post lethal infection, to the point where all mice were moribund, 80% of them were rescued by a single injection of this phage preparation. Based on the current results, a comprehensive study is needed to guide further research on the MRSA phage as a biocontrol for MRSA mastitis in dairy cows to replace or reduce the use of antibiotics in animal husbandry.